An 18- to 20-nm virus particle was isolated from the Olson strain of quail bronchitis, an avian adenovirus. On density gradient separation the small virions were primarily found at densities of 1.39 and 1.42 g/cm(3). The majority of the infectious particles were at the heavier density. The virus had a hexagonal outline and contained single-stranded deoxyribonucleic acid. It was resistant to heating at 56 C for more than an hour and was not inactivated by treatment with chloroform or low pH. Purified virus did not agglutinate erythrocytes of various avian and mammalian species. Replication of the small particles occurred either in chicken embryos or in cultures of embryo kidney cells coinfected with an adenovirus helper. Antigenically the virus was distinct from the adeno-associated viruses types 1, 2, 3, and 4. The virus is the avian equivalent of the adeno-associated viruses of primates and lower animals.